OBJECTIVETo compare the differences of the efficacy and adverse reaction of Oxaliplatin (L-OHP) treatment to oral squamous cell carcinoma (OSCC) at four different daily time points, and to analyze the characteristics of circadian rhythms.

METHODSSeventy-five nude mice were placed under 12h light and 12h dark cycles. Human OSCC cell line BcaCD885 was inoculated on the cheek of nude mice to establish a nude mice model of OSCC. After 3 weeks, mice were divided into 5 groups (4 experimental groups and 1 control group), with 15 in each group. L-OHP (17 mg x kg(-1)) was injected intravenously at 4 different time points during a period of 24 h, including 4 hours after lights on JHALO), 10 HALO, 16 HALO and 22 HALO for 4 experimental groups. The control group received normal saline of the same volume as that of L-OHP. The efficacy (tumor inhibition rate and survival time) and adverse reaction (body weight, white blood cell and perianal swelling) were observed after administration. The circadian rhythms of the efficacy and adverse reaction were examined by cosine analysis.

RESULTSL-OHP injected at 4, 16 and 22 HALO had great tumor inhibition rates, however, only 16 and 22 HALO groups significantly prolonged survival time of mice. The adverse reactions at 4 and 10 HALO were significantly severer than that of 16 and 22 HALO. Cosine analysis showed survival time, body weight and white blood cell counts had significant circadian rhythms. Mice received L-OHP at 14.88 HALO had the longest survival time.

CONCLUSIONThe time factor should be considered in L-OHP chronchemotherapy of patients with OSCC in order to increase the efficacy, decrease the adverse reaction of the drug and to improve the life quality of patients with OSCC.

Animals ; Carcinoma, Squamous Cell ; Circadian Rhythm ; Humans ; Mice ; Mice, Nude ; Mouth Neoplasms ; Organoplatinum Compounds

OBJECTIVEWe previously showed that introduction of a normal, neomycin-tagged human chromosome 8 reduced the metastatic capacity of C5F rat liver cancer cell line, which had high metastatic potential without affecting tumorigenicity, suggesting the presence of one or more metastasis suppressor genes encoded on human chromosome 8. We proceeded to define further the region harboring the metastasis suppressor gene(s) and to determine the random loss of human chromosome 8 by PCR amplification of sequence tag site (STS) markers.

METHODSThe national Center for Biotechnology Information (NCBI) databases were used as references of the relative genetic distances of the STS markers. C5F genomic DNA and A9/neo8 genomic DNA were used as negative and positive controls for chromosome 8 amplification, respectively. Genomic DNA was isolated and quantified from cultured hybrid clones (A9/C5F-1 and A9/C5F-2 microcell hybrid clones served as metastasis-unsuppressed groups; A9/C5F-4, A9/C5F-8 and A9/C5F-10 microcell hybrid clones served as metastasis suppressed groups). STS-PCR products were separated by electrophoresis through 2% agarose gel.

RESULTSMetastasis-suppressed microcell hybrid clones (A9/C5F-4, A9/C5F-8 and A9/C5F-10) conserved STS markers between D8S542 --> D8S1973 (8p21.1-23.1). In contrast, metastasis-unsuppressed clones (A9/C5F-1 and A9/C5F-2) lacked several markers in this region. In attempts to refine the region retained in the microcell suppressed clones, more densely spaced STS markers in the human chromosome 8p21.1-23.1 were used. We found that the metastasis-suppressed clones retained 18cM region between D8S542 and D8S1973 (8P21.1-23.1), where as the metastasis-unsuppressed clones lacked the region.

CONCLUSIONOur results suggest that a metastasis suppressor gene is located within the interval between D8S542 and D8S1973 on human chromosome 8p21.1-23.1.

Carcinoma, Hepatocellular ; genetics ; Cell Line ; Cell Line, Tumor ; Chromosome Mapping ; Chromosomes, Human, Pair 8 ; genetics ; Fibroblasts ; cytology ; Genes, Tumor Suppressor ; Humans ; In Situ Hybridization, Fluorescence ; Liver Neoplasms ; genetics ; Neoplasm Metastasis ; Sequence Tagged Sites

BACKGROUNDSynovium-derived stem cells (SDSCs) with higher chondrogenic potential are attracting considerable attention as a cell source for cartilage regeneration. We investigated the effect of bone morphogenetic protein 2 (BMP-2) on transforming growth factor beta3 (TGF-β3)-induced chondrogenesis of SDSCs isolated from human osteoarthritic synovium in a pellet culture system.

METHODSThe clonogenicity, stem cell marker expression and multi-differentiation potential of isolated SDSCs were determined by colony forming unit assay, flow cytometry and specific staining including alizarin red S, Oil red O and alcian blue staining, respectively. SDSCs pellet was cultured in chondrogenic medium with or without TGF-β3 or/and BMP-2. At day 21, the diameter and the weight of the pellets were measured. Chondrogenic differentiation of SDSCs was evaluated by Safranin O staining, immunohistochemical staining of collagen type II, sulfated glycosaminoglycan (sGAG) synthesis and mRNA expression of collagen type II, aggrecan, SOX9, link-protein, collagen type X and BMP receptor II.

RESULTSCells isolated under the optimized culturing density (10(4)/60 cm(2)) showed clonogenicity and multi-differentiation potential. These cells were positive (> 99%) for CD44, CD90, CD105 and negative (< 10%) for CD34 and CD71. SDSCs differentiated to a chondrocytic phenotype in chondrogenic medium containing TGF-β3 with or without BMP-2. Safranin O staining of the extracellular matrix was positive and the expression of collagen type II was detected. Cell pellets treated with TGF-β3 and BMP-2 were larger in diameter and weight, produced more sGAGs, and expressed higher levels of collagen type II and other chondrogenic markers, except COL10A1, than medium with TGF-β3 alone.

CONCLUSIONSSDSCs could be isolated from human osteoarthritic synovium. Supplementation with BMP-2 significantly promoted the in vitro TGF-β3-induced chondrogenic differentiation of SDSCs.

Aged ; Bone Morphogenetic Protein 2 ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chondrogenesis ; drug effects ; Female ; Humans ; Immunohistochemistry ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Middle Aged ; Synovial Membrane ; cytology ; Transforming Growth Factor beta3 ; pharmacology

OBJECTIVETo study the tumor cell killing function of T lymphocytes stimulated by dendritic cells (DC) and to analyze the differences of protein contents of exosomes in each type of cell.

METHODSThe exosomes of hepatic cell lines with high (P group) or low (F group) metastatic potentials were isolated by a process of four-step centrifugation and the collected exosomes were observed under an electron microscope (EM). The tumor cell killing experiment was performed by adding T lymphocytes activated by DC loaded with exosomes from corresponding P and F group cells and was studied using 3H-TdR experiments. The proteomic analysis was performed by surface-enhanced laser desorption/ ionization time of flight mass spectrometry (SELDI-TOF-MS ) on the exosomes of P and F group cells.

RESULTSThe density distribution and content of exosomes in the P group were not equal to those in the F group observed by EM. The CD80, CD86, MHC-I and MHC-II in the P group were 64.27+5.00, 44.89+10.11, 84.35+19.89 and 59.03+19.37, and those in the F group were 71.53+4.85, 50.01+9.50, 80.68+29.87 and 58.86+21.11, respectively (P>0.05, compared with the control group). The counts per minute value in the P group was 528.40+179.06 and 78.80+24.44 in the F group after being loaded with exosomes (P<0.01, compared with the control group). There were significant differences between the proteins in the exosomes of hepatic cancer cell lines with high or low metastatic potentials.

CONCLUSIONExosomes have potential values of application in immunotherapy and in biotherapy for recurrences and metastases of hepatic carcinomas.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Dendritic Cells ; immunology ; metabolism ; Exosomes ; Liver Neoplasms ; metabolism ; pathology ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; immunology ; metabolism

OBJECTIVETo investigate the effects of tamoxifen (TMX) combined with coenzyme Q10 (CoQ10) on idiopathic oligoasthenospermia.

METHODSA total of 183 patients with idiopathic oligoasthenospermia were randomly divided into a TMX + CoQ10 group (n = 63), a TMX group (n = 61) and a CoQ10 group (n = 59). At the end of 3 and 6 months of treatment, semen analyses and hormone tests were performed, and the results were compared with those obtained before the treatment.

RESULTSCompared with the pre-treatment results, the levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone (T) and sperm concentration were significantly elevated in the TMX + CoQ10 and TMX groups (P < 0.05), but showed no significant difference in the CoQ10 group (P > 0.05); sperm motility and morphologically normal sperm were increased significantly in the TMX + CoQ10 and CoQ10 groups (P < 0.05), and slightly in the TMX group but with no statistically significant difference (P > 0.05).

CONCLUSIONTamoxifen combined with CoQ10 can significantly improve sperm concentration, motility and morphology in patients with idiopathic oligoasthenospermia.

Adult ; Humans ; Male ; Oligospermia ; drug therapy ; Tamoxifen ; therapeutic use ; Treatment Outcome ; Ubiquinone ; analogs & derivatives ; therapeutic use ; Young Adult

OBJECTIVESTo study the relationship between the expression level of DLC-1 mRNA (located in 8p) and the invasion/metastasis of human hepatocellular carcinoma (HCC).

METHODSFifty-one surgical specimens of human HCC were divided into high-invasive and low invasive groups according to their clinicopathological features. DLC-1 mRNA expression was studied in the 51 HCC specimens as well as 5 different metastasis potential cell lines using real-time quantitative PCR (RQ-PCR).

RESULTSThe expression level of DLC-1 mRNA in HCC specimens with high invasiveness was significantly lower than that with low invasiveness (P < 0.05). The expression levels of DLC-1 mRNA were significantly different between non-metastatic (Hep3B and HepG2) and metastatic (MHCC97-H, MHCC97-L and HCCLM3) cell lines (P < 0.05). From MHCC97-L to HCCLM3, with an increase of invasiveness and metastatic potentials, the expression level of DLC-1 decreased correspondingly, and its expression level in HCCLM3 was significantly lower than that in MHCC97-L (P < 0.01).

CONCLUSIONThe expression of DLC-1 mRNA may play an important role in inhibiting the invasiveness and metastasis of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; GTPase-Activating Proteins ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Mutagenesis, Site-Directed ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Suppressor Proteins ; biosynthesis ; genetics

OBJECTIVETo develop an indirect immunofluorescence assay (IFA) for detection of IgG antibodies against new bunyavirus.

METHODSThe antigen slides were prepared with 5 new bunyavirus strains isolated using Africa green monkey kidney (Vero) cells. Specificity and sensitivity evaluation of IFA were carried out by optimizing working conditions of IFA. Using established IFA, serum samples from both acute and recovery phases were tested for 126 cases with fever thrombocytopenia and leukopenia syndrome in Xinyang, Henan province in 2007 - 2011. The results were compared with detections by RT-PCR.

RESULTSThe new bunyavirus stable immunofluorescence specific WZ69 strain was selected to prepare antigen slides of IFA. The optimum conditions of IFA were: optimum dilution for primary antibody (serum) and secondary antibody (isosulfocyanic acid fluorescence marked goat anti-human IgG antibody) was 1:40 and 1:150 respectively. The optimum dilution for Evans blue in secondary antibody was 1:20 000. Among the 126 patients, 96 paired serum specimens were tested positive to the new bunyavirus and 30 patients were tested negative to the virus. The positive rate of antibodies was 76.19%. There was no significant difference in results between IFA and RT-PCR (72.22% (91/126)) (P > 0.05).

CONCLUSIONThe IFA has high sensitivity and specificity with easy operation. It can be used in detecting the new bunyavirus infection in patients with fever, thrombocytopenia and leukopenia syndrome.

Animals ; Antibodies, Viral ; analysis ; immunology ; Bunyaviridae Infections ; diagnosis ; immunology ; Cercopithecus aethiops ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunoglobulin G ; analysis ; immunology ; Male ; Middle Aged ; Orthobunyavirus ; immunology ; isolation & purification ; Sensitivity and Specificity ; Vero Cells

In order to find new source of antifungal agents, eleven cultivable endophytic fungi were isolated from the roots,stems and leaves of Chelidonium majus by traditional method. Seven of them were identified as Colletotrichum(L1, L2, L3, S1, S3, S4, S5), and three of them were identified as Fusarium(R1,R2,R3) by morphological features and molecular biological technology. The antifungal activity test showed that all the tested fungi displayed some inhibitory activity against five common plant pathogens(C. gloeosporioides, Curvularia lunata, Pyricularia oryza, Alternaria alternate and A. brassicae), and their inhibition rate of some test items were over 60%. Among them, R1, S2, S3 and S4 were more potent than others. This study enriches the understanding of endophytes from Ch. majus and provides a basis for the study of new microbial fungicides.
Alternaria ; pathogenicity ; Antibiosis ; Ascomycota ; pathogenicity ; Chelidonium ; microbiology ; Colletotrichum ; chemistry ; isolation & purification ; Endophytes ; chemistry ; isolation & purification ; Fusarium ; chemistry ; isolation & purification

Autopsy of patients who died of infectious diseases is of significance for public health management. Few forensic autopsies have been performed since the outbreak of the corona virus disease 2019 （COVID-19） due to some limitations, thus forensic pathological examination failed to contribute to the prevention and treatment of infectious diseases. Virtual autopsy has unique advantages in the forensic examination of patients who died of infectious diseases. Accumulated virtual autopsy image data are of great value to the study of the pathological mechanism and diagnosis of COVID-19. This article reviews the relationship between imaging changes and pathology of the COVID-19 as well as the application of virtual autopsy in autopsy of patients who died of infectious diseases, in order to provide reference for performing virtual autopsy in the outbreak of COVID-19.
Autopsy ; Betacoronavirus ; COVID-19 ; Coronavirus Infections/pathology* ; Forensic Pathology ; Humans ; Pandemics ; Pneumonia, Viral/pathology* ; SARS-CoV-2

Objective: To explore the clinical characteristics and prognosis of the new coronavirus 2019-nCoV patients combined with cardiovascular disease （CVD). Methods: A retrospective analysis was performed on 112 COVID-19 patients with CVD admitted to the western district of Union Hospital in Wuhan, from January 20, 2020 to February 15, 2020. They were divided into critical group (ICU, n=16) and general group (n=96) according to the severity of the disease and patients were followed up to the clinical endpoint. The observation indicators included total blood count, C-reactive protein (CRP), arterial blood gas analysis, myocardial injury markers, coagulation function, liver and kidney function, electrolyte, procalcitonin (PCT), B-type natriuretic peptide (BNP), blood lipid, pulmonary CT and pathogen detection. Results: Compared with the general group, the lymphocyte count (0.74 (0.34, 0.94)×10⁹/L vs. 0.99 (0.71, 1.29)×10⁹/L, P=0.03) was extremely lower in the critical group, CRP (106.98 (81.57, 135.76) mg/L vs. 34.34 (9.55,76.54) mg/L, P<0.001) and PCT (0.20 (0.15,0.48) μg/L vs. 0.11 (0.06,0.20) μg/L, P<0.001) were significantly higher in the critical group. The BMI of the critical group was significantly higher than that of the general group (25.5 (23.0, 27.5) kg/m² vs. 22.0 (20.0, 24.0) kg/m²，P=0.003). Patients were further divided into non-survivor group (17, 15.18%) group and survivor group (95, 84.82%). Among the non-survivors, there were 88.24% (15/17) patients with BMI> 25.0 kg/m², which was significantly higher than that of survivors (18.95% (18/95), P<0.001). Compared with the survived patients, oxygenation index (130 (102, 415) vs. 434 (410, 444), P<0.001) was significantly lower and lactic acid (1.70 (1.30, 3.00) mmol/L vs. 1.20 (1.10, 1.60) mmol/L, P<0.001) was significantly higher in the non-survivors. There was no significant difference in the proportion of ACEI/ARB medication between the critical group and the general group or between non-survivors and survivors （all P>0.05). Conclusion: COVID-19 patients combined with CVD are associated with a higher risk of mortality. Critical patients are characterized with lower lymphocyte counts. Higher BMI are more often seen in critical patients and non-survivor. ACEI/ARB use does not affect the morbidity and mortality of COVID-19 combined with CVD. Aggravating causes of death include fulminant inflammation, lactic acid accumulation and thrombotic events.
Betacoronavirus ; COVID-19 ; Cardiovascular Diseases/therapy* ; Coronavirus Infections/complications* ; Humans ; Pandemics ; Pneumonia, Viral/complications* ; Prognosis ; Retrospective Studies ; SARS-CoV-2 ; Treatment Outcome